LOCUS       sequence-420115-        4814 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Rhizobium acidisoli LRD-modifying cytochrome P450 
            enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4814)
  AUTHORS   Lin GM, Voigt CA
  TITLE     Design of a redox-proficient Escherichia coli for screening 
            terpenoids and modifying cytochrome P450s
  JOURNAL   Nature Catalysis. 6, 1016–1029 (2023). 
            https://doi.org/10.1038/s41929-023-01049-5
REFERENCE   2  (bases 1 to 4814)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature 
            Catalysis. 6, 1016–1029 (2023). 
            https://doi.org/10.1038/s41929-023-01049-5"
COMMENT     Sequence Label: pCYP06
FEATURES             Location/Qualifiers
     source          1..4814
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(296..841)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(376..395)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(930..1745)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(930..1745)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1653..1672
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1737..1756
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    1922..1949
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt 
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     protein_bind    1980..2007
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt 
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     primer_bind     complement(4726..4745)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 actgactcgc tgcacgaggc agacctcagc gctagcggag tgtatactgg cttactatgt
       61 tggcactgat gagggtgtca gtgaagtgct tcatgtggca ggagaaaaaa ggctgcaccg
      121 gtgcgtcagc agaatatgtg atacaggata tattccgctt cctcgctcac tgactcgcta
      181 cgctcggtcg ttcgactgcg gcgagcggaa atggcttacg aacggggcgg agatttcctg
      241 gaagatgcca ggaagatact taacagggaa gtgagagggc cgcggcaaag ccgtttttcc
      301 ataggctccg cccccctgac aagcatcacg aaatctgacg ctcaaatcag tggtggcgaa
      361 acccgacagg actataaaga taccaggcgt ttccccctgg cggctccctc gtgcgctctc
      421 ctgttcctgc ctttcggttt accggtgtca ttccgctgtt atggccgcgt ttgtctcatt
      481 ccacgcctga cactcagttc cgggtaggca gttcgctcca agctggactg tatgcacgaa
      541 ccccccgttc agtccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
      601 gaaagacatg caaaagcacc actggcagca gccactggta attgatttag aggagttagt
      661 cttgaagtca tgcgccggtt aaggctaaac tgaaaggaca agttttggtg actgcgctcc
      721 tccaagccag ttacctcggt tcaaagagtt ggtagctcag agaaccttcg aaaaaccgcc
      781 ctgcaaggcg gttttttcgt tttcagagca agagattacg cgcagaccaa aacgatctca
      841 agaagatcat cttattaagg ggtctgacgc tcagtggaac gaaaaatcaa tctaaagtat
      901 atatgagtaa acttggtctg acagttacct tagaaaaact catcgagcat caaatgaaac
      961 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat
     1021 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg
     1081 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta
     1141 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagcttatgc
     1201 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca
     1261 tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg
     1321 ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca
     1381 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg
     1441 gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc
     1501 ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg
     1561 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat
     1621 cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa
     1681 tcagcatcca tgttggaatt taatcgcggc ctggagcaag acgtttcccg ttgaatatgg
     1741 ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt tcatgatgat
     1801 atatttttat cttgtgcaat gtacatcaga gattttgaga cacaaccaat tattgaaggc
     1861 ctccctaacg gggggccttt ttttgtttct ggtctcccaa gcgcttaacg atcgttggct
     1921 gaacaaacag acaatctggt ctgtttgtat tatggaaaat ttttctgtat aatagattca
     1981 acaaacagac aatctggtct gtttgtatta tcagcggtca acgcatgtgc tttgcgttct
     2041 gatgagacag tgatgtcgaa accgcctcta caaataattt tgtttaggtc agactataga
     2101 acccggaggt aaacaacaat ggacgtacaa gaaaccaagg cggcctgcca cgatgccttt
     2161 gcggagcttg caagtcccgc atgcattcaa gatccgtacc cttttatgcg ctggctgcaa
     2221 gaacacgatc cggttcatcg tgccgcatca ggcatatttc tgctgagccg tcatgctgat
     2281 atatattggg ccttcaaagc caccggtgat gcattccgcg gcccggcccc gagtgaattg
     2341 gcacgttatt ttccacgggc ggcctctagt ctgtctctga atttgttggc aagcactttg
     2401 gccatgaaag aaccgcccac ccatacgcgt ctgcgtcgcc tcatttcgcg agacttcacc
     2461 gtgggccaca ttgataactt acgaccgagc atagcgcgaa ttgttgctgc acggcttgat
     2521 ggtatggcgc ctgccctcga acgcggcgaa gcggtggatc tgcaccgtga attcgcgctc
     2581 gcattgccga tgctggtgtt cgccgaactg tttgggatgc cgcaggatga catttttgaa
     2641 cttagtgcta ttgtatccgc aatcctggag ggactctcac cgcacgcctc tgatccgcag
     2701 ttggcagcag ctgacgtcgc ctccgcgcgt gtgaaagcgt actttggcga cctcatcctg
     2761 cgcaaacgtg cggacccgtg ccgcgacatt gtatcgacgc tggtcggtgc tcattcggat
     2821 gacgccgaca cgttgtcaga cgctgaattg atttcgatgc tgtggggaat gctgctgggt
     2881 ggctttgcta ccactgcggc taccattgac catgcggtgc ttgccatgct ggcatatccc
     2941 gaagaacgtc attggctgca gggtgacgcg gcgggtgtcg aagcgtttgt cgaagaagtc
     3001 ctgcgttgcg aagcgccggc gatgttttct tcgattccgc gcatcgccca acgagatatc
     3061 gaattacgtg gcgtcgtgat ccctaaagac gcagatgttc gtgtattgat cgctgcgggt
     3121 aatcgtgatc cggatgcgtt cgcggaccca gaccggttcg acccggtgcg cttttatggg
     3181 actcgccctg gtatgagcaa cgatggcaaa attatgctca gcttcggtca tggtatccat
     3241 ttttgcttag gggcacagtt ggcccgtgtt cagttggcgg agagcttgcc acaaatacaa
     3301 gcacgcttcc caaccctggc cctggcggag cagccaacta gagaaccgag cgcttttctg
     3361 cgtacattcc gcgcactgcc ggttcgctta cacgcacagg ctgctgcaga ggtgagagta
     3421 gtggtggacc aagatctgtg cggcacgaca ggccagtgcg ttttgaccct gccaggtgca
     3481 tttcgtcaac gggaacctga tggagttgct gaagtctgca tggccactgt ccctcaggca
     3541 ttacatgccg ccgttcgtct ggcagagagc cagtgcccgg ttgccgctat tagagtaatt
     3601 gaatctgaag cgggcgataa ccactgtacc aatccgggtc ctacaccaag tcaagccgat
     3661 gcggagcgcc acgctgcgaa ggatctccgt aacccgggtg agcatgatgg caccatttaa
     3721 taaggatgag ctctacaaat aagcagaggt ggttgtgttg cgaaaaaaaa aaaaaaacac
     3781 cctaacgggt gttttttttt ttttggtgtc cccacgtgtg gcgctggaga ccgtccaatg
     3841 gcggcgcgcc atcgaatggt gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga
     3901 gagtcaattc agggtggtga atatgagccc gaaacgtcgt acccaggcag aacgtgcaat
     3961 ggaaacccag ggtaaactga ttgcagcagc actgggtgtt ctgcgtgaaa aaggttatgc
     4021 aggttttcgt attgcagatg ttccgggtgc agccggtgtt agccgtggtg cacagagcca
     4081 tcattttccg accaaactgg aactgctgct ggcaaccttt gaatggctgt atgagcagat
     4141 taccgaacgt agccgtgcac gtctggcaaa actgaaaccg gaagatgatg ttattcagca
     4201 gatgctggat gatgcagcag atttttttct ggatgatgat tttagcatcg gcctggatct
     4261 gattgttgca gcagatcgtg atccggcact gcgtgaaggt attctgcgta ccgttgaacg
     4321 taatcgtttt gttgttgaag atatgtggct gggtgtgctg gtgagccgtg gtctgagccg
     4381 tgatgatgcc gaagatattc tgtggctgat ttttaacagc gttcgtggtc tgacagttcg
     4441 tagcctgtgg cagaaagata aagaacgttt tgaacgtgtg cgtaatagca ccctggaaat
     4501 tgcacgtgaa cgttatgcaa aattcaaacg ttgataagga tcctaattgg taacgaatca
     4561 gacaattgac ggctcgaggg agtagcatag ggtttgcaga atccctgctt cgtccatttg
     4621 acaggcacat tatgcatcga tgataagctg tcaaacatga gcagatcctc tacgccggac
     4681 gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc tggcgcctat atcgccgaca
     4741 tcaccgatgg ggaagatcgg gctcgccact tcgggctcat gagcaaatat tttatctgag
     4801 gtgcttcctc gctc
//
